Off-Resonance TROSY (R1ρ − R1) for Quantitation of Fast Exchange Processes in Large Proteins
- 11 September 2003
- journal article
- research article
- Published by American Chemical Society (ACS) in Journal of the American Chemical Society
- Vol. 125 (40) , 12064-12065
- https://doi.org/10.1021/ja037101s
Abstract
Current solution NMR experiments for characterizing conformational exchange processes in large proteins are limited to exchange rates ca. 500−3000 s-1. A TROSY-based constant relaxation time (R1ρ− R1) experiment is designed to extend this capability to measure motion with rates up to 105 s-1 in large macromolecules. The experiment combines off-resonance spin-lock rf fields, which provide access to the faster time-scale dynamics, with TROSY coherence selection, which extends the molecular-weight range available for study. When implemented on the 53-kDa dimeric enzyme triosephosphate isomerase, the experiment yielded substantial gains in signal-to-noise (up to 60%) over current experiments at modest static magnetic fields (14.1 T). The TROSY (R1ρ−R1) experiment should therefore be of general utility for investigation of fast conformational exchange events in large proteins.Keywords
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